Tuesday, January 24, 2012

As not mentioned in Lombardi et al. 2009

January 22nd, 2010

Question: I’m confused about something. You say you can’t find it very easily by PCR, so you are culturing it. Then how did you find it for the Science paper?

Dr. Mikovits: Well, because I found it in 67%, by PCR, of the patients, but I looked at samples collected at several times. I just said it's like an EEG. So I got lucky [???] and I found it at a time when the patient was high [!!!]. They’ll come to the doctor when they’re sick [!!!], which might mean they’re replicating more white blood cells, and there’s more virus in their white blood cells.

Question: So you took like 10 samples from one patient?

Dr. Mikovits: Not that many, usually it was 4 [samples per patient]. The copy number could be as low as 5 or 10 copies per mil of blood, so there’s a statistic called a Poisson distribution, where you might find it one out of three times, so we went more than one of that and went 4 on most people when we found it.

So what is it Dr. Mikovits: Easy to find and everybody else is just too stupid to find it? Or it is somehow very hard and you have to be Lucky Mikovits and do several samples at several times to find it?

Yes, Dr. Mikovits is true scientific heroine.
Question: So the UK paper had studied patients, if they had looked at 4 or 5 per patient, do you think they might have found it?

Dr. Mikovits: They might have certainly found more, yeah.

Question: They essentially accused you of contamination.

Dr. Mikovits: Right. But why would I have contamination in my sick people and not my healthy people? [Not why, but how!] How would I do that? [Yeah, how? XMRV VP62 plasmid and 5AZA maybe?] I did it in three different labs. I did it in Cleveland Clinic. When I first came to the Institute, we didn’t have a lab. So everything we drew from around the world, I sent to Frank’s office. They processed it. I went there, I put it in the microarray or did whatever I was going to do, and we worked there for a couple of weeks, you know, while we were building our lab and it got started. [Suddenly it is not "I found …" but "They processed …"???]

From the way we did the study, there’s just NO possible way there was contamination. And that’s what the reviewers concluded. [The reviewers were blind mole people] And the phylogenetic analysis was one of the things they asked for (they asked for 3 things after the initial submission). The phylogenetic analysis proved two things. It wasn’t a mouse virus, it’s a human virus; it wasn’t a contaminant from a lab. We never do mouse work, but it wasn’t a contaminant from mouse feces or something in the lab [Except XMRV VP62 plasmid maybe], and it clearly was a new branch in the tree, a human virus. And our virus wasn’t exactly the same as the prostate cancer virus. It’s still XMRV, because it’s 99% similar. But that’s enough to show it’s not a contaminant. [SAY WHAT???] One of the things you have to use to get a good PCR is at least 750 nanograms of DNA. They have no idea how much DNA was there. And they quantitated 3-9 out of 186? Sure they found a band of globins, but globins are in every single cell, so again you’re making an unfair comparison of what you’re saying you see. And then you amplify it for fewer cycles than what would really push the envelope.

We’ve also done - to show no mouse contamination with the CDC, and Bill Switzer - after he saw the results in the paper before it became published. He said, “I have an assay that’ll show it’s a mouse contamination. It’s a very sensitive, very specific PCR. Will you do it?” I said, “Sure, send it to me.” [because it won't pick up XMRV VP62 plasmid] We did it on all 100 and not a one. Not one cell line in our lab. He’s found it in a couple in his lab, but we didn’t find any. Perfectly controlled. He said, “Congratulations, it’s not a mouse contaminant.” [because it is a plasmid contaminant] So there’s little else we can do, except wait for the rest of the community. It’s there. [no, it isn't] And the prostate cancer people didn’t say: “oh you didn’t find it in 500 people; it must not have anything to do with prostate cancer." Because it just didn’t have anything to do with that population. [Look! Over there!]

And again, Norbert Bannert is a high-quality scientist. [unlike myself] As soon as he saw the paper, he called me and asked for the reagents. Because he’s gonna go back and look to see if it really is there, and help us find some answers. [he thought you were a scientist] He’s also looking at a CFS group. So it just depends on what you really want to find. We weren’t biased in our study. You know, I’m a cancer cell biologist, and we aren’t biased. [Yeah, right.] I had to work really hard to get most of the people - not Frank and company - the National Cancer Institute (NCI) didn't know what CFS was. You know, fortunately, our scientists can to some level do what they want, as long as it’s along with the mission. I remember one gentleman, high level NCI official, said “tell ‘em to get over it.” Again that’s the credit to Annette and the formation of the WPI - we got a grant where we literally named XMRV within my first six months in 2007, because of the juxtaposition of seeing that paper in prostate cancer, right when we met.

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